Engineering of daidzein 3’-hydroxylase P450 enzyme into catalytically self-sufficient cytochrome P450
<p>Abstract</p> <p>A cytochrome P450 (CYP) enzyme, 3’-daidzein hydroxylase, CYP105D7 (3’-DH), responsible for daidzein hydroxylation at the 3’-position, was recently reported. CYP105D7 (3’-DH) is a class I type of CYP that requires electrons provided through electron transfer prote...
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| Format: | Article |
| Language: | English |
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BMC
2012-06-01
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| Series: | Microbial Cell Factories |
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| Online Access: | http://www.microbialcellfactories.com/content/11/1/81 |
| _version_ | 1818118650955038720 |
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| author | Choi Kwon-Young Jung EunOk Jung Da-Hye An Byeo-Ri Pandey Bishnu Yun Hyungdon Sung Changmin Park Hyung-Yeon Kim Byung-Gee |
| author_facet | Choi Kwon-Young Jung EunOk Jung Da-Hye An Byeo-Ri Pandey Bishnu Yun Hyungdon Sung Changmin Park Hyung-Yeon Kim Byung-Gee |
| author_sort | Choi Kwon-Young |
| collection | DOAJ |
| description | <p>Abstract</p> <p>A cytochrome P450 (CYP) enzyme, 3’-daidzein hydroxylase, CYP105D7 (3’-DH), responsible for daidzein hydroxylation at the 3’-position, was recently reported. CYP105D7 (3’-DH) is a class I type of CYP that requires electrons provided through electron transfer proteins such as ferredoxin and ferredoxin reductase. Presently, we constructed an artificial CYP in order to develop a reaction host for the production of a hydroxylated product. Fusion-mediated construction with the reductase domain from self-sufficient CYP102D1 was done to increase electron transfer efficiency and coupling with the oxidative process. An artificial self-sufficient daidzein hydroxylase (3’-ASDH) displayed distinct spectral properties of both flavoprotein and CYP. The fusion enzyme catalyzed hydroxylation of daidzein more efficiently, with a <it>k</it><sub>cat</sub>/K<sub>m</sub> value of 16.8 μM<sup>-1</sup> min<sup>-1</sup>, which was about 24-fold higher than that of the 3’-DH-camA/B reconstituted enzyme. Finally, a recombinant <it>Streptomyces avermitilis</it> host for the expression of 3’-ASDH and production of the hydroxylated product was developed. The conversion that was attained (34.6%) was 5.2-fold higher than that of the wild-type.</p> |
| first_indexed | 2024-12-11T04:57:41Z |
| format | Article |
| id | doaj.art-1f493b8455944e84a3e335bf7d7214f2 |
| institution | Directory Open Access Journal |
| issn | 1475-2859 |
| language | English |
| last_indexed | 2024-12-11T04:57:41Z |
| publishDate | 2012-06-01 |
| publisher | BMC |
| record_format | Article |
| series | Microbial Cell Factories |
| spelling | doaj.art-1f493b8455944e84a3e335bf7d7214f22022-12-22T01:20:13ZengBMCMicrobial Cell Factories1475-28592012-06-011118110.1186/1475-2859-11-81Engineering of daidzein 3’-hydroxylase P450 enzyme into catalytically self-sufficient cytochrome P450Choi Kwon-YoungJung EunOkJung Da-HyeAn Byeo-RiPandey BishnuYun HyungdonSung ChangminPark Hyung-YeonKim Byung-Gee<p>Abstract</p> <p>A cytochrome P450 (CYP) enzyme, 3’-daidzein hydroxylase, CYP105D7 (3’-DH), responsible for daidzein hydroxylation at the 3’-position, was recently reported. CYP105D7 (3’-DH) is a class I type of CYP that requires electrons provided through electron transfer proteins such as ferredoxin and ferredoxin reductase. Presently, we constructed an artificial CYP in order to develop a reaction host for the production of a hydroxylated product. Fusion-mediated construction with the reductase domain from self-sufficient CYP102D1 was done to increase electron transfer efficiency and coupling with the oxidative process. An artificial self-sufficient daidzein hydroxylase (3’-ASDH) displayed distinct spectral properties of both flavoprotein and CYP. The fusion enzyme catalyzed hydroxylation of daidzein more efficiently, with a <it>k</it><sub>cat</sub>/K<sub>m</sub> value of 16.8 μM<sup>-1</sup> min<sup>-1</sup>, which was about 24-fold higher than that of the 3’-DH-camA/B reconstituted enzyme. Finally, a recombinant <it>Streptomyces avermitilis</it> host for the expression of 3’-ASDH and production of the hydroxylated product was developed. The conversion that was attained (34.6%) was 5.2-fold higher than that of the wild-type.</p>http://www.microbialcellfactories.com/content/11/1/81Cytochrome P450 monooxygenasesDaidzein 3’-hydroxylaseSelf-sufficient P450CYP102D1 |
| spellingShingle | Choi Kwon-Young Jung EunOk Jung Da-Hye An Byeo-Ri Pandey Bishnu Yun Hyungdon Sung Changmin Park Hyung-Yeon Kim Byung-Gee Engineering of daidzein 3’-hydroxylase P450 enzyme into catalytically self-sufficient cytochrome P450 Microbial Cell Factories Cytochrome P450 monooxygenases Daidzein 3’-hydroxylase Self-sufficient P450 CYP102D1 |
| title | Engineering of daidzein 3’-hydroxylase P450 enzyme into catalytically self-sufficient cytochrome P450 |
| title_full | Engineering of daidzein 3’-hydroxylase P450 enzyme into catalytically self-sufficient cytochrome P450 |
| title_fullStr | Engineering of daidzein 3’-hydroxylase P450 enzyme into catalytically self-sufficient cytochrome P450 |
| title_full_unstemmed | Engineering of daidzein 3’-hydroxylase P450 enzyme into catalytically self-sufficient cytochrome P450 |
| title_short | Engineering of daidzein 3’-hydroxylase P450 enzyme into catalytically self-sufficient cytochrome P450 |
| title_sort | engineering of daidzein 3 hydroxylase p450 enzyme into catalytically self sufficient cytochrome p450 |
| topic | Cytochrome P450 monooxygenases Daidzein 3’-hydroxylase Self-sufficient P450 CYP102D1 |
| url | http://www.microbialcellfactories.com/content/11/1/81 |
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